摘要
目的 建立乳酸杆菌及嗜热链球菌的PFGE分子分型方法,并对北京市售酸奶中分离的乳酸杆菌及嗜热链球菌进行分子分型.方法 选取ApaⅠ、NotⅠ、sfiⅠ、XbaⅠ和SmaⅠ共5种PFGE分析中常用的限制性内切酶,对从北京市售酸奶中分离到的52株乳酸杆菌、嗜热链球菌以及相应的标准菌株进行酶切,优化PFGE限制性内切酶种类及电泳条件,并用优化出的实验条件对菌株进行分子分型,同时进行聚类分析,与生化鉴定及16s rRNA基因鉴定结果进行对比分析.结果 限制性内切酶NotⅠ对保加利亚乳酸杆菌、发酵乳酸杆菌和德氏乳酸杆菌的酶切效果较好,而限制性内切酶Apa Ⅰ对嗜热链球菌、嗜酸乳酸杆菌及干酪乳酸杆菌的酶切效果较好.24株保加利亚乳酸杆菌被分为8个PFGE型,15株嗜热链球菌被分为8个PFGE型,7株嗜酸乳酸杆菌被分为3个PFGE型,2株德氏乳酸杆菌分属于2个不同的PFGE型.结论 建立的PFGE方法分析结果与生化鉴定及16s rRNA基因鉴定结果高度符合,所建方法适用于乳酸杆菌及嗜热链球菌的分子分型.
Objective To develop a pulsed field gel electrophoresis （PFGE） method for molecular typing of Lactobacillus and Streptococcus thermophilus （S.thermophilus） and to apply it in identification and characterization of both bacteria isolated from yoghourt collected from Beijing supermarket.Methods The five most useful restriction enzymes includingApa Ⅰ,Not ],Sfi Ⅰ,Xba Ⅰ and Sma Ⅰ were chosen to cut DNA of 52 strains of Lactobacillus,S.thermophilus as well as associated standard bacteria strains.The endonucleases and electrophoresis conditions for PFGE analysis were optimized and applied in molecular typing of Lactobacillus and S.thermophilus isolates.Cluster analysis based on the PFGE data was conducted.The identification results of PFGE were compared with those obtained in biochemical and 16s ribosomal RNA PCR identification tests.Results Not Ⅰ was suitable for L.bulgaricus,L.fermentum and L.delbrueckii digestion.While Apa Ⅰ was an appropriate endonuclease for S.thermophilus,L.acidophilus and L.casei digestion.The results of molecular typing indicated that 24 strains of L.bulgaricus and 15 strains of S.thermophilus were grouped into 8 types by PFGE method,respectively.While 7 strains of L.acidophilus were grouped into 3 types and 2 strains of L.delbrueckii were grouped into 2 different PFGE types.Conclusion The results of PFGE analysis are in compliance with those of 16s rRNA PCR and biochemical identification.The PFGE method developed in this study is suitable for molecular characterization of both Lactobacillus and S.thermophilus.
出处
《中华预防医学杂志》
CAS
CSCD
北大核心
2011年第12期1086-1089,共4页
Chinese Journal of Preventive Medicine
基金
“十一五”国家科技支撑计划（2009BADB9B00）
关键词
乳酸杆菌属
链球菌
嗜热
细菌分型技术
电泳
凝胶
脉冲场
Lactobacillus
Streptococcus thermophilus
Bacterial typing techniques
Electrophoresis, gel ,pulsed-field
